Direct cDNA Kit
Description
YB Direct RT-PCR kit provides a fast and simple procedure for preparing first-strand cDNA directly from cultured cells without RNA purification. RNA in the lysate can be directly converted to cDNA and subsequently analyzed using the PCR reagents included in the kit or real-time RT-PCR kits purchased elsewhere. The whole process can be completed in less than 1.5 hours. Because no removal of genomic DNA is included in the protocol, users need to design primers from the exon-exon junctions of the target mRNA to allow detection of RNA only.
Features
    • Rapid :
      prepare cell lysate from cultured cell within 10 min and utilized directly for RT-PCR
    • Simple :
      from cells to cDNA in only 2 steps and under 0.5 hour
    • Sensitive :
      detection of even low-abundance transcripts
    • Time saving :
      easy parallel processing of several samples
    • One-tube system :
      rapid, sensitive and reproducible analysis of RNA with minimal risk of sample contamination
    • High thermostability :
      Deoxy+ MMLV reverse transcriptase is used
  • Convenient format :
    premixed solutions for use in RT-qPCR
Applications
  • Generation of cDNA products with high fidelity for cloning and sequencing
  • Gene expression analysis
  • Analysis of a large number of differentially treated cultures
  • Using in real-time PCR
Quality Control
The performance of YB Direct RT-PCR Kit is tested in an RT reaction using human embryonic kidney cell lysate with primer d(T)18. The sensitivity of the kit is verified by the detection of B2M transcript after 35 cycles. The length of cDNA achieved is verified by detection of a 248 bp by DNA agarose gel and DNA sequencing.
Results

 
 
Specification
Store at 

 

# FYT507-50P (50 preps) 

Wash Buffer 25 ml 
Cell Lysis Buffer 1.25 ml×2
Deoxy+ HiSpec Reverse Transcriptase 100 μl
2× RT Buffer 500 ul
10 μM d(T)20 50 μl
Nuclease-free Water 1 ml

# FYT507-100P (100 preps) 

Wash Buffer 50 ml  
Cell Lysis Buffer 1.25 ml ×4
Deoxy+ HiSpec Reverse Transcriptase 200 μl
2× RT Buffer 1 ml
10 μM d(T)20 100 μl
Nuclease-free Water 2 ml